The DC generation core facility will consist of two components located at the Baylor Institute for Immunology Research and the Rockefeller University. The cores' primary responsibility will be prepare GMP grade DCs from stage III and stage IV melanoma patients for the clinical trials proposed in projects 1-3. These include DCs derived from CD34+ hematopoietic progenitors [CD34-DC], and monocyte-derived DCs [MDC]. The core will follow standard operating procedures that have been established at each institution for the generation of each type of DC. The cores have successfully generated high quality cells for in vivo studies in healthy volunteers and stage IV melanoma patients. The cores will also provide 2 additional functions: [a] adapt alternative approaches to mature DCs for clinical use, and [b] adapt novel approaches for loading DCs with dying melanoma cells for vaccination purposes. New approaches to mature DCs will be investigated in projects 2 and 3, including the use of poly I:C, TNF alpha and PGE2 vs. standard stimuli e.g. monocyte-conditioned medium. Cross-presentation of antigens from dying cells represent novel approaches to present a broad repertoire of tumor antigens including both CD4 and CD8 epitopes. The core will customize procedures to generate mature GMP grade DCs that have been fed apoptotic cells. In summary the DC core will provide a unique opportunity to optimize and standardize protocols to generate DCs for studies of human tumor immunology and immunotherapy.